Phosphorylation of RPT6 controls its ability to bind DNA and regulate gene expression in the hippocampus during memory formation. Phosphorylation of RPT6 controls its ability to bind DNA and regulate gene expression in the hippocampus during memory formation

We used RNA-sequencing along with novel genetic approaches and biochemical, molecular, and behavioral assays to test the hypothesis that pRPT6-S120 functions independently of the proteasome to bind DNA and regulate gene expression during memory formation. RNA-sequencing following siRNA-mediated knockdown of free RPT6 revealed 46 gene targets in the dorsal hippocampus of male rats following fear conditioning, where RPT6 was involved in transcriptional activation and repression. Through CRISPR-dCas9-mediated artificial placement of RPT6 at a target gene promoter, we found that RPT6 DNA binding alone was sufficient for altering gene expression following learning. Further, CRISPR-dCas13-mediated conversion of S120 to glycine on RPT6 revealed that phosphorylation at S120 is necessary for RPT6 to bind DNA and properly regulate transcription during memory formation. Together, we reveal a novel function for phosphorylation of RPT6 in controlling gene transcription during memory formation. Overall design: Male Sprague-Dawley rats were injected into the dorsal hippocampus CA1 region with Accell siRNA against Psmc5 or control (Scr-siRNA). Five days later animals were trained to contextual fear conditioning and the CA1 region (dorsal) was dissected, RNA collected via columns and used for RNA-seq analysis